• 대한바이러스학회지
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• 제28권2호
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• pp.129-138
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• 1998

Bovine growth hormone (bGH) gene was expressed in an insect Spodoptera frugiperda cell line using a Baculovirus, Hyphantria cunea nuclear polyhedrosis virus (HcNPV). The bGH gene in pbGH plasmid was sequenced and amplified by PCR technique with two primers containing NcoI sites. The bGH gene consisted of 654 bp (217 amino acid residues), the 5'-untranslated region of the cloned bGH cDNA contains 56 bp, and the 3'-untranslated region contains 145 bp and two pallindromic regions. The amplified bGH gene DNA fragment (654 bp) was inserted into the NcoI site of the pHcEVII vector, which was named pHcbGH. The pHcbGH transfer vector DNA and the wild type HcNPV DNA were cotransfected into S. frugiperda cells to construct a recombinant virus. Eight recombinant viruses were selected and named HcbGH. One clone, HcbGH-4-1 showed largest plaque size, therefore the recombinant virus was further studied. The multiplication pattern of the recombinant HcbGH-4-1 was similar to that of the wild type HcNPV. The bGH gene DNA in the HcbGH-4-1 recombinant was confirmed by Southern blot hybridization. The amount of the bGH (217 amino acid residues, 21 kDa) produced in S. frugiperda cells infected with the HcbGH-4-1 recombinant was approximately 5.5 ng per ml ($10^6$ cells) by radioimmunoassay.

• Journal of mucopolysaccharidosis and rare diseases
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• 제1권1호
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• pp.19-22
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• 2015

In Growth Hormone (GH) therapy, suboptimal adherence is a common problem, reaching up to 82%, and there is a need for interventions to improve adherence and to maximize patients' growth potential eventually. Current studies have demonstrated the association between the rate of non-adherence and reduced height velocity. In order to maximize patients' potential to grow, an auto-injecting/recording device, such as $easypod^{TM}$, may help improve adherence and optimize the treatment effects of GH therapy. The use of $easypod^{TM}$ has contributed to high adherence rates: 87.5% and 93% in Bozzola et al.'s study and the $Easypod^{TM}$ Connect Observational study (ECOS), respectively. Improvement of adherence by $easypod^{TM}$ may lead to higher growth rates of patients receiving GH therapy. Additionally, patients' positive acceptability of $easypod^{TM}$ suggests $easypod^{TM}$ is a preferred device by patients for better adherence.

• 한국의류산업학회지
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• 제5권1호
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• pp.77-81
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• 2003

The purpose of this study was to investigate the effect of multi-functional fabric on EEG and growth hormone (GH) during sleep and quality of sleep with the 9 young female athletes. The subjects participated in separated experimental procedure; sleeping in multi-functional fabric wear (experimental group) and cotton wear (control group) for 450min. During the night (22:00-05:30), we recorded the changes of nocturnal polysomnographic sleep recording and GH were measured every 60min. The results show that there are significant differences in percentage of stage 1, 2 and slow wave sleep (SWS) between two groups(S1, p<.05; S2, SWS, p<.01). The SWS percentage of experimental group is 1.89 times higher than control group. The changes of GH secretion varied depending on two experimental procedures. The peak of GH secretion in experimental group is more than controls by 2.4time (p<.001). The quality of sleep in experimetal group is significantly higher than control (p<.01). These results suggest muti-functional fabric wear is effective in inducing the deep sleep and increasing GH and quality of sleep.

• BMB Reports
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• 제34권6호
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• pp.502-508
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• 2001

A cDNA, encoding the human growth hormone (hGH), was synthesized based on the known 191 amino acid sequence. Its codon usage was optimized for a high level expression in Escherichia coli. Unique restriction sites were incorporated throughout the gene to facilitate mutagenesis in further studies. To minimize an initiation translation problem, a 624-bp cassette that contained a ribosome binding site and a start codon were fused to the hGH-coding sequence that was flanked between the EcoRI and HindIII sites. The whole fragment was synthesized by an overlapped extension of eight long synthetic oligonucleotides. The four-short duplexes of DNA, which were first formed by annealing and filling-in with a Klenow fragment, were assembled to form a complete hGH gene. The hGH was cloned and expressed successfully using a pET17b plasmid that contained the T7 promoter. Recombinant hGH yielded as much as 20% of the total cellular proteins. However, the majority of the protein was in the form of insoluble inclusion bodies. N-terminal amino acid sequencing also showed that the hGH produced in E. coli contained formyl-methionine. This study provides a useful model for synthesis of the gene of interest and production of recombinant proteins in E. coli.

• Journal of mucopolysaccharidosis and rare diseases
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• 제1권2호
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• pp.54-59
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• 2015

Purpose: Growth hormone (GH) therapy substantially improves several cognitive functions in PWS. However, the molecular mechanisms underlying the beneficial effects of GH on cognition remain unclear in PWS. In this study, we investigated the effects of recombinant human GH on the gene expression of GABAB receptor subunits and GH/insulin-like growth factor (IGF) axis genes in the brain regions of PWS-mimicking mice (Snord116del). Methods: Snord116del mice were injected subcutaneously with 1.0 mg/kg GH or saline, once daily for 7 days. The collected brain tissues were analyzed for mRNA content using quantitative PCR (qPCR) in the cerebellum, hippocampus, and cerebral cortex. Results: GH increased the mRNA expression level of the $GABA_{B1}$ receptor subunit ($GABA_{BR1}$) and IGF-1R in the cerebellum. Furthermore, a significant positive correlation was found between the level of $GABA_{BR1}$ mRNA and the expression of the IGF-1R transcript. GH also induced an increase in the mRNA expression of IGF-2 and IGF-2R in the cerebellum. Conclusion: These data indicate that GH may provide beneficial effects on cognitive function through its influences on the expression of $GABA_{BR1}$ and GH/IGF-1 axis genes in PWS patients.

• Journal of mucopolysaccharidosis and rare diseases
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• 제3권2호
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• pp.41-43
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• 2017

Prader-Willi syndrome (PWS) is a genetic disorder that is considered, especially on child, to cause poor feeding, hypotonia, failure to thrive, developmental delay and hypogonadism which is known to affect between 1 in 10,000 and 30,000 people. The children with PWS are viewed as affected by growth hormone (GH) insufficiency, although the exact mechanisms of GH deficiency are not fully understood. However, the benefits of GH treatment in children with PWS are well established. Myers, et al. (2006), Grugni, et al. (2016) indicated its positive effects on linear growth, body composition, motor function, respiratory function and psychomotor development. Despite of its effectiveness and advantages had been well known and proven in many other studies, there is only one recombinant GH product that is approved for PWS in Korea, $Genotropin^{(R)}$, till now. A phase III clinical study of GH treatment with $Eutropin^{TM}$, in 34 Korean PWS children is in progress, which is expected to have comparable effects and safety profile with the active control by assessing auxological changes such as height standard deviation score, body composition changes such as lean body mass and percent body fat, motor and cognitive development using Bayley scale, and safety profiles.

• Clinical and Experimental Pediatrics
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• 제62권7호
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• pp.274-280
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• 2019

Purpose: To analyze the growth response to growth hormone (GH) therapy in prepubertal patients with Noonan syndrome (NS) harboring different genetic mutations. Methods: Twenty-three patients with prepubertal NS treated at Pusan National University Children's Hospital between March 2009 and July 2017 were enrolled. According to the disease-causing genes identified, the patients with NS were divided into 4 groups. Three groups were positive for mutations of the PTPN11, RAF1, and SOS1 genes. The five genes undetected (FGU) group was negative for PTPN11, RAF1, SOS1, KRAS, and BRAF gene mutations. The influence of genotype was retrospectively analyzed by comparing the growth parameters after GH therapy. Results: The mean chronological age at the start of GH treatment was $5.85{\pm}2.67years$. At the beginning of the GH treatment, the height standard deviation score (SDS), growth velocity (GV), and lower levels of insulin-like growth factor-1 (IGF)-1 levels were not statistically different among the groups. All the 23 NS patients had significantly increased height SDS and serum IGF-1 level during the 3 years of treatment. GV was highest during the first year of treatment. During the 3 years of GH therapy, the PTPN11, RAF1, and SOS1 groups showed less improvement in height SDS, IGF-1 SDS, and GV, and less increase in bone age-to-chronological age ratio than the FGU group. Conclusion: The 3-year GH therapy in the 23 prepubertal patients with NS was effective in improving height SDS, GV, and serum IGF-1 levels. The FGU group showed a better response to recombinant human GH therapy than the PTPN11, RAF1, and SOS1 groups.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.251-256
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• 2011

Human growth hormone (hGH), one of the most important hormones in medicine, is secreted from anterior pituitary gland. Its broad physiological function includes body growth, cell regeneration, increasement of muscle volume, bone density, body fat reduction, and so on. Due to the wide range of therapeutic effects, the hGH produced from E. coli has been commercialized already. In this study, we asked whether it is possible to produce recombinant hGH efficiently from various cultured mammalia cells. To meet this purpose, we chose a retrovirus vector system for transfer and expression of the hGH gene in various mammalian cells. Analyses of RT-PCR, ELISA, and Western blot to determine expression of the hGH gene showed the highest production of the hGH was determined from chicken embronic fibroblast (CEF) cells with the concentration of 8.58 ${\mu}g$/ml. The biological activity of the hGH was similar to the commercially available counterpart. These results suggest that mass production of hGH is possible not only in the E. coli but also in the various mammalian cells.

• Reproductive and Developmental Biology
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• 제35권4호
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• pp.519-525
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• 2011

Growth hormone (GH) is obligatory for growth and development. But, there is controversy on the GH effect about reproductive processes of sexual differentiation, pubertal maturation, gonadal steroidogenesis, gametogenesis and ovulation. This study was conducted to investigate the effect of GH on estrus, ovulation and embryo implantation. The results obtained were as follows. GH stimulated to increase estrus rate (p<0.05), pregnancy rate (p<0.05), and total fetus number in mice treated for superovulation. Also, the correlation between GH and steroids, E2 and P4, at peri-estrus stage/ peri-ovulation stage/ peri-implantation stage of the superovulation-induced mice was examined. Consequently, GH co-injected with PMSG especially increased P4 level (p<0.05) at peri-estrus stage of superovulationinduced mice. In conclusion, GH co-treatment in superovulation system boosted the rate of estrus, pregnancy and total fetus by increasing progesterone level at peri-estrus stage of superovulation-induced mice.

• 한국발생생물학회지:발생과생식
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• 제2권2호
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• pp.189-196
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• 1998

Growth Hormone Releasing Hormone (GHRH)은 척추동물의 시상하부로부터 합성, 분비되어 시상하부-뇌하수체간의 문맥계를 통해 뇌하수체 전엽에 작용하여 Growth Hormone (GH)의 분비를 촉진한다. 시상하부에서 발현되는 일부 Releasing Hormone 들이 여러 시상하부외 조직에서도 검출되고 조직특이적인 기능을 수행한다는 사실이 여러 연구자들에 의해 밝혀졌다. 이러한 사실들을 배경으로 본 연구자는 GHRH가 흰쥐의 뇌하수체 전엽과 뇌하수체로부터 유래된 종양세포주들에서 발현될 가능성을 조사하였다. GHRH 펩타이드와 mRNA의 존재와 구조를 규명하기 위하여 뇌하수체와 배양 세포를 사용하여 GHRH immunocytochemistry, 방사면역측정법, GHRH PCR과 RNase protection assay를 시행하였다. Immunocytochemistry의 결과 gonadotrope (대형)와 somat-olactotrope (중간형)로 추정되는 세포들에서 GHRH 염색이 나타났고, Somatolactotrope성 종양세포인 GH3 cell 추출물에서 immunoreactive GHRH가 방사면역측정법으로 검출되었다. 3'rapid amplification of cDNA end (3'-RACE)를 시행한 결과, 흰쥐 뇌하수체에 GHRH transcript가 존재하고, 그 3'end 부분이 다른 조직내의 GHRH와 동일함을 확인하였다. GHRH RT-PCR에서도 뇌하수체와 종양세포주들인 $\alpha$T3 cell (gonadotrope성)과 GH3 cell에서 예상 산물들이 증폭되었다. RNase protection assay를 시행한 결과 난소절제에 의해 뇌하수체내 GHRH 유전자 발현이 증가됨을 확인하였다. 이상의 결과는 GHRH가 뇌하수체 전엽의 gonadotrope와 somatotrope에서 발현되고, paracrine 또는 autocrine조절물질로 작용하여 GH 분비 외에도 뇌하수체 전엽 세포들의 분화와 분열등에 관여함을 시사한다.